【摘要】 目的探讨低氧诱导因子1(HIF1)在肾微血管内皮细胞(HGMECs)缺氧再复氧损伤中的免疫调节作用。方法建立HGMECs缺氧再复氧模型,应用特异性脯氨酰羟化酶抑制剂3,4DHB和环氧酶2阻断药物NS398预处理HGMECs,设对照、缺氧复氧、3,4DHB和NS398四组。采用逆转录聚合酶链反应、免疫印记和酶联免疫吸附试验等方法检测各组细胞HIF1α、血红素氧合酶1(HO1)的mRNA和蛋白的表达及上清中可溶性细胞间黏附因子1(sICAM1)的水平;将各组HGMECs与异体淋巴细胞混合培养(MELR),检测淋巴细胞增殖程度和MELR上清中白细胞介素2(IL2)的表达水平。结果与对照组相比,HGMECs经缺氧复氧损伤后HIF1α蛋白和HO1mRNA表达水平显著增高(P<0.01),上清中sICAM1(14.55±1.13vs7.90±1.61)ng、异体淋巴细胞增殖率(0.191±0.053vs0.069±0.032)和MELR上清中IL2(40.0±5.0vs18.0±3.0)pg水平均有显著增高(P<0.01)。与缺氧复氧组比较,3,4DHB干预组HIF1α蛋白和HO1mRNA表达水平显著增高(P<0.05),而淋巴细胞增殖率(0.079±0.038,P<0.01)及IL2(25.7±6.1pg,P<0.01)和sICAM1(8.71±1.32ng,P<0.01)水平则显著下调。与缺氧复氧组比较,NS398干预组HIF1α蛋白和HO1mRNA表达水平显著减低(P<0.01),而淋巴细胞增殖率(0.268±0.079)及IL2(51.0±11.0)pg和sICAM1(20.33±3.05)ng表达水平则显著增高(P<0.05)。结论缺氧复氧会对HGMECs造成免疫学损伤,HIF1α稳定表达可起到保护性作用。更多还原

【Abstract】 Objective To explore the influence of hypoxia inducible factor-1 (HIF-1) on the immunofunction of human glomerular microvascular endothelial cells (HGMECs) with hypoxia-reoxygenation (HR).Methods Model of HGMECs induced by HR was established to mimic ischemia/reperfusion injury.Prolyl 4-hydroxylase inhibitor 3,4-DHB and cyclooxygenase enzymes-2 (COX-2) blocker NS-398 were used to intervene the expression of HIF-1 and heme oxygenase-1 (HO-1).Proliferative rate of lymphocytes stimulated by the HGMECs was measured by MTT test,while the supernatant levels of IL-2 and ICAM-1 were detected by ELISA.Results As compared with HGMECSs with HR,the expression of HIF-1α protein and HO-1 mRNA was significantly increased in HGMECs pretreated with 3,4-DHB (P<0.05).While the supernatant levels of IL-2 [(25.7± 6.1) vs (40.0± 5.0)pg, P< 0.01],ICAM-1 [(8.71± 1.32) vs (14.55± 1.13)ng, P< 0.01] and the lymphocyte proliferative rate (0.079± 0.038 vs 0.191± 0.053, P= 0.003) were significantly decreased.On the contrary,the expression of HIF-1α protein and HO-1 mRNA was markedly decreased in the HGMECs pretreated with NS-398 as compared with the HGMECs with HR (P< 0.05).Conclusion The overexpression of HIF-1α may have protective immunofunction on the HGMECs with HR.更多还原